Powder: -20°C for 3 years | In solvent: -80°C for 1 year
AZ3146 是一种选择性 Mps1 抑制剂,IC50为 35 nM。
规格 | 价格/CNY | 货期 | 数量 | |
---|---|---|---|---|
1 mg | ¥ 255 | 现货 | ||
2 mg | ¥ 356 | 现货 | ||
5 mg | ¥ 567 | 现货 | ||
10 mg | ¥ 912 | 现货 | ||
25 mg | ¥ 1,680 | 现货 | ||
50 mg | ¥ 2,860 | 现货 | ||
100 mg | ¥ 4,230 | 现货 | ||
1 mL * 10 mM (in DMSO) | ¥ 725 | 现货 |
产品描述 | AZ3146 is a selective Mps1 inhibitor with IC50 of ~35 nM. |
靶点活性 | Mps1:35 nM |
体内活性 | AZ3146不影响Aurora B和BubR1的有丝分裂特异性磷酸化形式。HeLa细胞用诺考达唑和2 μM AZ3146处理,仅短暂延迟有丝分裂,随后重新复制它们的基因组,表明AZ3146能够覆盖SAC。值得注意的是,90% AZ3146处理的HeLa细胞发生异常有丝分裂,~50%进入分裂后期的细胞没有调整它们的染色体,~30%完成有丝分裂的细胞没有进行明显的染色体分离。AZ3146也抑制FAK,JNK1,JNK2 和 Kit。AZ3146明显抑制细胞中Mps1的磷酸化作用。AZ3146也会抑制已经建立的SAC信号,如从诺考达唑阻断释放后,AZ3146明显加速有丝分裂结束。AZ3146对Mad2的着丝粒定位有显著影响,将其降至~15%,但是它对Mad1的作用不明显,其水平保持在~60%。进入有丝分裂期前,Mps1被AZ3146抑制,随后Mad1和Mad2对着丝粒的聚集被阻止。然而,如果Mps1在有丝分裂进入后被AZ3146抑制,则Mad1-C-Mad2核心复合物仍然与着丝粒结合,但O-Mad2不能聚集到核心。在其他未受到干扰的有丝分裂中,AZ3146使完成有丝分裂的时间从对照组的90分钟减少到32分钟。 |
激酶实验 | His-tagged human Mps1Cat encoding amino acids 510-857 is generated. For kinase assays, 500 ng is added to buffer (25 mM Tris-HCl, pH 7.4, 100 mM NaCl, 50 μg/mL BSA, 0.1 mM EGTA, 0.1% β-mercaptoethanol, 10 mM MgCl2, and 0.5 μg/mL myelin basic protein), AZ3146, and 100 μM γ-[32P]ATP (2 μCi/assay). Reactions are incubated at 30°C for 20 min, spotted onto P81 paper, washed in 0.5% phosphoric acid, and immersed in acetone. Phosphate incorporation is determined by scintillation counting. For immunoprecipitation kinase assays, HeLa cells are treated with nocodazole for 14 h, mitotic cells isolated, washed in PBS, and lysed for 30 min in 50 mM Tris-HCl, pH 7.4, 100 mM NaCl, 0.5% NP-40, 5 mM EDTA, 5 mM EGTA, 40 mM β-glycerophosphate, 0.2 mM PMSF, 1 mM DTT, 1 mM sodium orthovanadate, 20 mM sodium fluoride, 1 μM okadaic acid, and complete EDTA-free protease inhibitor cocktail. Full-length Mps1 is immunoprecipitated. Purified complexes are washed with lysis buffer containing 100 mM NaCl and assayed as described for the recombinant protein. To quantify 32P incorporation, reactions are stopped with SDS sample buffer and separated by SDS-PAGE followed by phosphorimaging. The plate is analyzed using a phosphorimager using AIDA software. To assess the specificity of AZ3146, a single-point screen is carried using kinase profiling service. 50 kinases are selected and assayed with 1 μM AZ3146[1]. |
细胞实验 | AZ3146 is disolved in DMSO (100 mM) and diluted into 100 μM, 10 μM, 1 μM and 0.1 μM sequentially with DMEM containing 10% FBS before use[2]. The TTK inhibitor AZ3146 is disolved in DMSO at a concentration in 100 mM and diluted into 100 μM, 10 μM, 1 μM and 0.1 μM sequentially with DMEM containing 10% FBS before use. In vitrocytotoxicity assays are performed. HCC cells are plated into 96-well plates at the density of 3×103 per well. AZ3146 is added in the indicated concentrations the next day. The inhibitor treated cells are cultured and tested at a 24-hour intervals for 3-4 days using CCK-8[2]. |
别名 | AZ 3146 |
分子量 | 452.55 |
分子式 | C24H32N6O3 |
CAS No. | 1124329-14-1 |
Powder: -20°C for 3 years | In solvent: -80°C for 1 year
DMSO: 11.3 mg/mL (25 mM)
Ethanol: 33.9 mg/mL (75 mM)
可选溶剂 | 浓度 体积 质量 | 1 mg | 5 mg | 10 mg | 25 mg |
DMSO / Ethanol | 1 mM | 2.2097 mL | 11.0485 mL | 22.097 mL | 55.2425 mL |
5 mM | 0.4419 mL | 2.2097 mL | 4.4194 mL | 11.0485 mL | |
10 mM | 0.221 mL | 1.1049 mL | 2.2097 mL | 5.5243 mL | |
20 mM | 0.1105 mL | 0.5524 mL | 1.1049 mL | 2.7621 mL | |
Ethanol | 50 mM | 0.0442 mL | 0.221 mL | 0.4419 mL | 1.1049 mL |
对于不同动物的给药剂量换算,您也可以参考 更多...
请在以下方框中输入您的动物实验信息后点击计算,可以得到母液配置方法和体内配方的制备方法: 比如您的给药剂量是10 mg/kg,每只动物体重20 g,给药体积100 μL,一共给药动物10 只,您使用的配方为5% DMSO+30% PEG300+5% Tween 80+60% ddH2O。那么您的工作液浓度为2 mg/mL。
母液配置方法:2 mg 药物溶于 50 μL DMSO (母液浓度为 40 mg/mL), 如您需要配置的浓度超过该产品的溶解度,请先与我们联系。
体内配方的制备方法:取 50 μL DMSO 主液,加入 300 μL PEG300, 混匀澄清,再加 50 μL Tween 80,混匀澄清,再加 600 μL ddH2O, 混匀澄清。
您可能有的问题的答案可以在抑制剂处理说明中找到,包括如何准备库存溶液,如何存储产品,以及基于细胞的分析和动物实验需要特别注意的问题。
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